Abstract
Pyrobaculum arsenaticum is a hyperthermophilic archaeon that thrives at 95°C. This strain encodes a putative GH31 intracellular α-glucosidase (Pars_2044, PyAG) in its genome. The recombinant PyAG (rPyAG) was optimally expressed in Escherichia coli at 37°C for 4 h after IPTG induction. The purified rPyAG is a homotetrameric α-glucosidase that exhibited highly thermostable properties. Maximum p-nitrophenyl-α-D-glucopyranoside (pNPG) hydrolysis activity was observed at 90°C and pH 5.0. The enzyme mainly recognized the non-reducing end of the substrate, releasing the glucose unit. rPyAG also had broad substrate specificity, cleaving maltose (α-1,4-linkage), kojibiose (α-1,2-linkage), and nigerose (α-1,3-linkage) with similar efficiency. Based on these results, rPyAG can be used to modify health-relevant sugar conjugates linked by α-1,2- or α-1,3-bonds.
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Okuyama M, Saburi W, Mori H, Kimura A. α-Glucosidases and α-1,4-glucan lyases: Structures, functions, and physiological actions. Cell Mol. Life Sci. 73: 2727–2751 (2016)
Stam MR, Danchin EG, Rancurel C, Coutinho PM, Henrissat B. Dividing the large glycoside hydrolase family 13 into subfamilies: Towards improved functional annotations of α-amylase-related proteins. Protein Eng. Des. Sel. 19: 555–562 (2006)
Park I, Lee H, Cha J. Glycoconjugates synthesized via transglycosylation by a thermostable α-glucosidase from Thermoplasma acidophilum and its glycosynthase mutant. Biotechnol. Lett. 36: 789–796 (2014)
Castanys-Munoz E, Martin MJ, Prieto PA. 2’-fucosyllactose: An abundant, genetically determined soluble glycan present in human milk. Nutr. Rev. 71: 773–789 (2013)
Feinberg LF, Srikanth R, Vachet RW, Holden JF. Constraints on anaerobic respiration in the hyperthermophilic archaea Pyrobaculum islandicum and Pyrobaculum aerophilum. Appl. Environ. Microb. 74: 396–402 (2008)
Jung JH, Seo DH, Holden JF, Park CS. Identification and characterization of an archaeal kojibiose catabolic pathway in the hyperthermophilic Pyrococcus sp. strain ST04. J. Bacteriol. 196: 1122–1131 (2014)
Li D, Park JT, Li X, Kim S, Lee S, Shim JH, Park SH, Cha J, Lee BH, Kim JW, Park KH. Overexpression and characterization of an extremely thermostable maltogenic amylase, with an optimal temperature of 100 degrees C, from the hyperthermophilic archaeon Staphylothermus marinus. New Biotechnol. 27: 300–307 (2010)
Jung JH, Seo DH, Holden JF, Park CS. Maltose-forming α-amylase from the hyperthermophilic archaeon Pyrococcus sp. ST04. Appl. Microbiol. Biot. 98: 2121–2131 (2014)
Jeon EJ, Jung JH, Seo DH, Jung DH, Holden JF, Park CS. Bioinformatic and biochemical analysis of a novel maltose-forming α-amylase of the GH57 family in the hyperthermophilic archaeon Thermococcus sp. CL1. Enzyme Microb. Tech. 60: 9–15 (2014)
Li X, Li D, Park KH. An extremely thermostable amylopullulanase from Staphylothermus marinus displays both pullulan-and cyclodextrin-degrading activities. Appl. Microbiol. Biot. 97: 5359–5369 (2013)
Huber R, Sacher M, Vollmann A, Huber H, Rose D. Respiration of arsenate and selenate by hyperthermophilic archaea. Syst. Appl. Microbiol. 23: 305–314 (2000)
Ernst HA, Lo Leggio L, Willemoes M, Leonard G, Blum P, Larsen S. Structure of the Sulfolobus solfataricus α-glucosidase: Implications for domain conservation and substrate recognition in GH31. J. Mol. Biol. 358: 1106–1124 (2006)
Park JE, Park SH, Woo JY, Hwang HS, Cha J, Lee H. Enzymatic properties of a thermostable α-glucosidase from acidothermophilic crenarchaeon Sulfolobus tokodaii strain 7. J. Microbiol. Biotechn. 23: 56–63 (2013)
Tan K, Tesar C, Wilton R, Keigher L, Babnigg G, Joachimiak A. Novel α-glucosidase from human gut microbiome: Substrate specificities and their switch. FASEB J. 24: 3939–3949 (2010)
Song JM, An YJ, Kang MH, Lee YH, Cha SS. Cultivation at 6-10 degrees C is an effective strategy to overcome the insolubility of recombinant proteins in Escherichia coli. Protein Expres. Purif. 82: 297–301 (2012)
Koma D, Sawai T, Harayama S, Kino K. Overexpression of the genes from thermophiles in Escherichia coli by high-temperature cultivation. Appl. Microbiol. Biot. 73: 172–180 (2006)
Jeon H, Lee H, Byun D, Choi H, Shim JH. Molecular cloning, characterization, and application of a novel thermostable α-glucosidase from the hyperthermophilic archaeon Pyrobaculum aerophilum strain IM2. Food Sci. Biotechnol. 24: 175–182 (2015)
Seo SH, Choi KH, Hwang S, Kim J, Park CS, Rho JR, Cha J. Characterization of the catalytic and kinetic properties of a thermostable Thermoplasma acidophilum α-glucosidase and its transglucosylation reaction with arbutin. J. Mol. Catal. B-Enzym. 72: 305–312 (2011)
Kim S, Lee SB. Soluble expression of archaeal proteins in Escherichia coli by using fusion-partners. Protein Expres. Purif. 62: 116–119 (2008)
Kang MS, Okuyama M, Mori H, Kimura A. The first α-1,3-glucosidase from bacterial origin belonging to glycoside hydrolase family 31. Biochimie 91: 1434–1442 (2009)
Saburi W, Okuyama M, Kumagai Y, Kimura A, Mori H. Biochemical properties and substrate recognition mechanism of GH31 α-glucosidase from Bacillus sp. AHU 2001 with broad substrate specificity. Biochimie 108: 140–148 (2015)
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Jung, JH., Seo, DH., Holden, J.F. et al. Broad substrate specificity of a hyperthermophilic α-glucosidase from Pyrobaculum arsenaticum . Food Sci Biotechnol 25, 1665–1669 (2016). https://doi.org/10.1007/s10068-016-0256-7
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DOI: https://doi.org/10.1007/s10068-016-0256-7